Enhanced in vivo selection of bone marrow cells by retroviral-mediated coexpression of mutant O6-methylguanine-DNA-methyltransferase and HOXB4.

2.50
Hdl Handle:
http://hdl.handle.net/10541/78379
Title:
Enhanced in vivo selection of bone marrow cells by retroviral-mediated coexpression of mutant O6-methylguanine-DNA-methyltransferase and HOXB4.
Authors:
Milsom, Michael D; Woolford, Lorna B; Margison, Geoffrey P; Humphries, R Keith; Fairbairn, Leslie J
Abstract:
To attain therapeutic levels of gene-modified hematopoietic stem cells, it may be necessary in the majority of disorders to provide an in vivo selective advantage that facilitates the expansion of their numbers. A popular strategy to achieve in vivo selection has been to employ drug selection while coexpressing a transgene that conveys chemoresistance, such as O6-methylguanine-DNA-methyltransferase (MGMT). An alternate approach is to confer an enhanced proliferative potential upon gene-modified hematopoietic stem cells through the delivery of the homeobox transcription factor HOXB4. By developing a novel tricistronic retroviral vector, we have facilitated the simultaneous coexpression of a mutant version of MGMT and HOXB4 in retrovirally transduced bone marrow. Using an in vivo competitive repopulation assay, we demonstrate that primary bone marrow cells containing this construct show enhanced reconstitution following transplant and improved selection subsequent to chemotherapeutic challenge in comparison to cells expressing either HOXB4 or MGMT alone. This selection advantage was evident even when HOXB4/MGMT-coexpressing cells were infused along with a large excess of unmodified cells. We propose that this selection cassette may facilitate the in vivo expansion of gene-modified hematopoietic stem cells at a level in excess of previous strategies.
Affiliation:
Gene Therapy, Paterson Institute for Cancer Research, Christie Hospital NHS Trust, Manchester M20 4BX, UK.
Citation:
Enhanced in vivo selection of bone marrow cells by retroviral-mediated coexpression of mutant O6-methylguanine-DNA-methyltransferase and HOXB4. 2004, 10 (5):862-73 Mol. Ther.
Journal:
Molecular Therapy
Issue Date:
Nov-2004
URI:
http://hdl.handle.net/10541/78379
DOI:
10.1016/j.ymthe.2004.07.019
PubMed ID:
15509504
Type:
Article
Language:
en
ISSN:
1525-0016
Appears in Collections:
All Paterson Institute for Cancer Research

Full metadata record

DC FieldValue Language
dc.contributor.authorMilsom, Michael D-
dc.contributor.authorWoolford, Lorna B-
dc.contributor.authorMargison, Geoffrey P-
dc.contributor.authorHumphries, R Keith-
dc.contributor.authorFairbairn, Leslie J-
dc.date.accessioned2009-08-24T14:24:11Z-
dc.date.available2009-08-24T14:24:11Z-
dc.date.issued2004-11-
dc.identifier.citationEnhanced in vivo selection of bone marrow cells by retroviral-mediated coexpression of mutant O6-methylguanine-DNA-methyltransferase and HOXB4. 2004, 10 (5):862-73 Mol. Ther.en
dc.identifier.issn1525-0016-
dc.identifier.pmid15509504-
dc.identifier.doi10.1016/j.ymthe.2004.07.019-
dc.identifier.urihttp://hdl.handle.net/10541/78379-
dc.description.abstractTo attain therapeutic levels of gene-modified hematopoietic stem cells, it may be necessary in the majority of disorders to provide an in vivo selective advantage that facilitates the expansion of their numbers. A popular strategy to achieve in vivo selection has been to employ drug selection while coexpressing a transgene that conveys chemoresistance, such as O6-methylguanine-DNA-methyltransferase (MGMT). An alternate approach is to confer an enhanced proliferative potential upon gene-modified hematopoietic stem cells through the delivery of the homeobox transcription factor HOXB4. By developing a novel tricistronic retroviral vector, we have facilitated the simultaneous coexpression of a mutant version of MGMT and HOXB4 in retrovirally transduced bone marrow. Using an in vivo competitive repopulation assay, we demonstrate that primary bone marrow cells containing this construct show enhanced reconstitution following transplant and improved selection subsequent to chemotherapeutic challenge in comparison to cells expressing either HOXB4 or MGMT alone. This selection advantage was evident even when HOXB4/MGMT-coexpressing cells were infused along with a large excess of unmodified cells. We propose that this selection cassette may facilitate the in vivo expansion of gene-modified hematopoietic stem cells at a level in excess of previous strategies.en
dc.language.isoenen
dc.subjectHaematopoietic Stem Cell Transplantationen
dc.subject.meshAnimals-
dc.subject.meshBone Marrow Cells-
dc.subject.meshCell Line-
dc.subject.meshColony-Forming Units Assay-
dc.subject.meshDacarbazine-
dc.subject.meshDeoxyguanosine-
dc.subject.meshGenetic Vectors-
dc.subject.meshGreen Fluorescent Proteins-
dc.subject.meshHematopoietic Stem Cell Transplantation-
dc.subject.meshHomeodomain Proteins-
dc.subject.meshMice-
dc.subject.meshMutation-
dc.subject.meshO(6)-Methylguanine-DNA Methyltransferase-
dc.subject.meshRetroviridae-
dc.subject.meshTranscription Factors-
dc.titleEnhanced in vivo selection of bone marrow cells by retroviral-mediated coexpression of mutant O6-methylguanine-DNA-methyltransferase and HOXB4.en
dc.typeArticleen
dc.contributor.departmentGene Therapy, Paterson Institute for Cancer Research, Christie Hospital NHS Trust, Manchester M20 4BX, UK.en
dc.identifier.journalMolecular Therapyen

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